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Literature summary for 7.2.1.1 extracted from

  • Juarez, O.; Neehaul, Y.; Turk, E.; Chahboun, N.; DeMicco, J.M.; Hellwig, P.; Barquera, B.
    The role of glycine residues 140 and 141 of subunit B in the functional ubiquinone binding site of the Na+-pumping NADH:quinone oxidoreductase from Vibrio cholerae (2012), J. Biol. Chem., 287, 25678-25685.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 7.2.1.1

Protein Variants

Protein Variants Comment Organism
G140A subunit NqrB. Site-directed mutagenesis, the mutant shows highly reduced activity and reduced inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide compared to the wild-type enzyme Vibrio cholerae serotype O1
G140A sodium-pumping activity of the mutant is not affected under partial turnover conditions. Mutation affects exclusively the binding of ubiquinone Vibrio cholerae serotype O1
G140L subunit NqrB, site-directed mutagenesis. The mutant shows highly reduced activity compared to the wild-type enzyme and no inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide in contrast to the wild-type enzyme Vibrio cholerae serotype O1
G140L mutant shows nonsaturating behavior with up to 50 microM ubiquinone Vibrio cholerae serotype O1
G141A subunit NqrB, site-directed mutagenesis. The mutant shows reduced activity compared to the wild-type enzyme and no inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide in contrast to the wild-type enzyme Vibrio cholerae serotype O1
G141A overall structure of the enzyme is not disturbed by the mutation. Mutation specifically affects the ubiquinone binding site. 6fold increase in Km value Vibrio cholerae serotype O1
G141L 3fold increase in Km value Vibrio cholerae serotype O1
G141L subunit NqrB, site-directed mutagenesis. The mutant shows reduced activity compared to the wild-type enzyme and no inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide in contrast to the wild-type enzyme Vibrio cholerae serotype O1
G141V site-directed mutagenesis, the mutant shows reduced activity and reduced inhibition by 2-n-heptyl-4-hydroxyquinoline N-oxide compared to the wild-type enzyme Vibrio cholerae serotype O1
G141V 9fold increase in Km value Vibrio cholerae serotype O1

Inhibitors

Inhibitors Comment Organism Structure
2-n-heptyl-4-hydroxyquinoline N-oxide
-
 Vibrio cholerae serotype O1

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics Vibrio cholerae serotype O1
0.0025
-
 ubiquinone-1 wild-type, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0025
-
 ubiquinone wild-type enzyme, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0088
-
 ubiquinone NqrB, mutant G141L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0088
-
 ubiquinone-1 NqrB, mutant G141L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.015
-
 ubiquinone NqrB, mutant G141A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.015
-
 ubiquinone-1 NqrB, mutant G141A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0234
-
 ubiquinone NqrB, mutant G141V, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0234
-
 ubiquinone-1 NqrB, mutant G141V, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.1
-
 ubiquinone above, mutant NqrB-G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.1
-
 ubiquinone-1 mutant G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.1
-
 ubiquinone-1 mutant G140L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
NADH + H+ + ubiquinone + n Na+/in Vibrio cholerae serotype O1
-
 NAD+ + ubiquinol + n Na+/out
-
 ?
NADH + H+ + ubiquinone + n Na+/in Vibrio cholerae serotype O1 ATCC 39541
-
 NAD+ + ubiquinol + n Na+/out
-
 ?

Organism

Organism UniProt Comment Textmining
Vibrio cholerae serotype O1 A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4 A5F5X1: subunit NqrA, A5F5X0: subunit NqrB, A5F5Y7: subunit NqrC, A5F5Y6: subunit NqrD; A5F5Y3: subunit NqrE, A5F5Y4: subunit NqrF. The enzyme consists of six subunits encoded by the NQR operon.
-
Vibrio cholerae serotype O1 ATCC 39541 A5F5X1 and A5F5X0 and A5F5Y7 and A5F5Y6 and A5F5Y3 and A5F5Y4 A5F5X1: subunit NqrA, A5F5X0: subunit NqrB, A5F5Y7: subunit NqrC, A5F5Y6: subunit NqrD; A5F5Y3: subunit NqrE, A5F5Y4: subunit NqrF. The enzyme consists of six subunits encoded by the NQR operon.
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
NADH + H+ + ubiquinone + n Na+/in
-
 Vibrio cholerae serotype O1 NAD+ + ubiquinol + n Na+/out
-
 ?
NADH + H+ + ubiquinone + n Na+/in NqrB-G140 and -G141 are critical for the binding and reaction of Na+-NQR with its electron acceptor, ubiquinone Vibrio cholerae serotype O1 NAD+ + ubiquinol + n Na+/out
-
 ?
NADH + H+ + ubiquinone + n Na+/in
-
 Vibrio cholerae serotype O1 ATCC 39541 NAD+ + ubiquinol + n Na+/out
-
 ?
NADH + H+ + ubiquinone + n Na+/in NqrB-G140 and -G141 are critical for the binding and reaction of Na+-NQR with its electron acceptor, ubiquinone Vibrio cholerae serotype O1 ATCC 39541 NAD+ + ubiquinol + n Na+/out
-
 ?
NADH + H+ + ubiquinone-1 + n Na+/in
-
 Vibrio cholerae serotype O1 NAD+ + ubiquinol-1 + n Na+/out
-
 ?
NADH + H+ + ubiquinone-1 + n Na+/in
-
 Vibrio cholerae serotype O1 ATCC 39541 NAD+ + ubiquinol-1 + n Na+/out
-
 ?

Synonyms

Synonyms Comment Organism
Na+-NQR
-
 Vibrio cholerae serotype O1
Na+-pumping NADH:quinone oxidoreductase
-
 Vibrio cholerae serotype O1
NqrB
-
 Vibrio cholerae serotype O1

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
200
-
 ubiquinone mutant G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
200
-
 ubiquinone-1 mutant G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
200
-
 ubiquinone-1 mutant G140L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
388
-
 ubiquinone mutant G141V, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
388
-
 ubiquinone-1 mutant G141V, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
425
-
 ubiquinone mutant G141A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
425
-
 ubiquinone-1 mutant G141A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
463
-
 ubiquinone mutant G141L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
463
-
 ubiquinone-1 mutant G141L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone-1 wild-type, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone wild-type enzyme, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Vibrio cholerae serotype O1

Cofactor

Cofactor Comment Organism Structure
FAD non-covalently bound in the NqrF subunit Vibrio cholerae serotype O1
FMN two covalently bound in subunits NqrB und NqrC Vibrio cholerae serotype O1
NADH
-
 Vibrio cholerae serotype O1
riboflavin one non-covalently bound in the subunit NqrB Vibrio cholerae serotype O1

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
 additional information inhibition kinetics Vibrio cholerae serotype O1
0.0005
-
 2-n-heptyl-4-hydroxyquinoline N-oxide wild-type enzyme, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0005
-
 2-n-heptyl-4-hydroxyquinoline N-oxide wild-type, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0012
-
 2-n-heptyl-4-hydroxyquinoline N-oxide mutant G141V, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0075
-
 2-n-heptyl-4-hydroxyquinoline N-oxide above, mutant G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
0.0075
-
 2-n-heptyl-4-hydroxyquinoline N-oxide mutant G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1

General Information

General Information Comment Organism
malfunction glycine 140 and glycine 141 of the NqrB subunit in the functional binding of ubiquinone. Mutations at these residues alter the affinity of the enzyme for ubiquinol. Mutations in residue NqrBG140 almost completely abolished the electron transfer to ubiquinone Vibrio cholerae serotype O1
metabolism the enzyme is the main entrance for electrons into the respiratory chain of many marine and pathogenic bacteria. The enzyme accepts electrons from NADH and donates them to ubiquinone, and the free energy released by this redox reaction is used to create an electrochemical gradient of sodium across the cell membrane Vibrio cholerae serotype O1
additional information subunit NqrB residues G140 and G141 are critical for the binding and reaction of Na+-NQR with its electron acceptor, ubiquinone Vibrio cholerae serotype O1

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
525
-
 ubiquinone-1 wild-type, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone-1 mutant G140A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone-1 mutant G140L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone-1 mutant G141A, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone-1 mutant G141L, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1
525
-
 ubiquinone-1 mutant G141V, pH 8.0, temperature not specified in the publication Vibrio cholerae serotype O1